Figure 4

BNIP3 is a mediator of OGD-induced hippocampal neuronal death. (A) Representative western blot showing the expression of BNIP3. (B) Quantification of the expression of BNIP3. Data were presented as fold of the control group. *p < 0.05 vs. control group, n = 3. (C) Forced expression of BNIP3 by transfection with EGFP-C3-rBNIP3 plasmid in normal cultured hippocampal neurons induced cell death. EGFP-C3 plasmid was transfected to neurons as a control. *p < 0.05, n = 21-29. (D-E) Inhibition efficiency of BNIP3-miRNA on BNIP3 expression in HEK293 cells co-transfected with indicated plasmids. *p < 0.05 vs. neg-miRNA transfected control, n = 3. (F) Cultured neurons were transfected with BNIP3-miRNA or neg-miRNA 48 h before OGD. Cell death of positive transfected neurons (green) was assessed by nuclear DAPI staining (blue) after OGD. Neurons pretreated with BNIP3-miRNA showed less pyknosis 12 h after reoxygenation compared with neg-miRNA treated neurons. (G) Quantification of experiments described in (F). Pretreatment with BNIP3-miRNA significantly reduced cell death compared with neg-miRNA treated controls. *p < 0.05 vs. control group, n = 21-26.